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. 2017:2017:2810565.
doi: 10.1155/2017/2810565. Epub 2017 Aug 15.

Stereospecific Assay of (R)- and (S)-Goitrin in Commercial Formulation of Radix Isatidis by Reversed Phase High-Performance Liquid Chromatography

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Stereospecific Assay of (R)- and (S)-Goitrin in Commercial Formulation of Radix Isatidis by Reversed Phase High-Performance Liquid Chromatography

Lixing Nie et al. J Anal Methods Chem. 2017.

Abstract

Radix isatidis (Banlangen) is a widely used traditional Chinese medicine for treating fever and removing toxic heat. Pharmacological studies have indicated that (R)-goitrin (epigoitrin) is one of the main constituents accounting for its antiviral activity, while (S)-goitrin (goitrin) is known as an antithyroid factor. To better control the quality of radix isatidis and its formulations, it is imperative to enantiomerically determine the contents of R- and S-goitrin. In this study, an enantioselective method based on reversed phase chromatography was developed for the assay of (R, S)-goitrin enantiomers. Optimum separation was obtained on an S-Chiral A column (4.6 mm × 250 mm, 5 μm) using methanol/water (30 : 70, v/v) as the mobile phase. After validation, the method was applied to quantify the enantiomers in Banlangen granules, which is the most prescribed commercial formulation of radix isatidis. Compared to enantioselective resolution approaches based on normal phase chromatography, the new method could be conveniently performed using regular reversed phase high-performance liquid chromatography (RP-HPLC) equipment and was found to be more suitable for the enantioselective quality control of water-extracted and soluble medicines.

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Figures

Figure 1
Figure 1
Chemical structures of (R, S)-goitrin.
Figure 2
Figure 2
Effect of (a) methanol concentration, (b) flow rate, and (c) column temperature on the resolution of (R, S)-goitrin enantiomers. Experimental conditions: S-Chiral A column (4.6 mm × 250 mm, 5 μm), methanol/water, 245 nm, and injection volume of 10 μL.
Figure 3
Figure 3
Effect of (a) methanol concentration, (b) flow rate, and (c) column temperature on the retention time of (R)-goitrin (▲) and (S)-goitrin (■). Experimental conditions: S-Chiral A column (4.6 mm × 250 mm, 5 μm), methanol/water, 245 nm, and injection volume of 10 μL.
Figure 4
Figure 4
Typical chromatograms obtained for the blank reference, standard solution containing racemic (R, S)-goitrin, and commercial granules. Experimental conditions: S-Chiral A column (4.6 mm × 250 mm, 5 μm), methanol/water (30 : 70, v/v), 0.4 mL/min, 35°C, 245 nm, and injection volume of 10 μL.

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